Body composition changes and their relationship with obstructive sleep apnoea symptoms, severity: The Sleeping Well Trial.

BACKGROUND & AIMS: Obstructive sleep apnoea (OSA) and obesity share a complex bi-directional relationship as location of body fat and changes in regional body composition may be more important for OSA improvement than changes in total body weight only. The aim of this study was to evaluate the impact of a 6-month weight loss intervention for adults newly diagnosed with moderate-severe OSA and obesity on regional body composition. The secondary aims evaluated the relationship between changes in OSA symptoms and severity and anthropometry and regional body composition during the first 12-months after commencing CPAP and explored differences in outcomes between males and females. METHODS: Participants (n = 59) received CPAP overnight at home alongside a 6-month modified fasting intervention with 12-months follow up. Regional body composition was measured by Dual X-ray absorptiometry, (DXA) and anthropometry before and after the lifestyle intervention. OSA severity was measured using the apnoea hypopnea index via overnight polysomnography and OSA symptoms were measured using the Epworth Sleepiness scale. RESULTS: Forty-seven adults (74% male) had complete measures available with a mean age of 50.0 y (SD 11.0) and BMI 34.1 kg/m2 (SD 5.0). Following the intervention average fat mass changed by -5.27 kg (5.36), p < 0.001) and visceral adipose tissue (-0.63 kg (0.67), p < 0.001) significantly decreased in males only with a maintenance of fat-free mass (mean -0.41 kg (1.80), p = 0.18). Females (n = 12) had significant decreases in waist circumference (mean -3.36 cm (3.18) p < 0.01), android lean (-0.12 kg (0.04), p < 0.05) and android total mass (-0.28 kg (0.39), p < 0.05) only. Regional body composition changes in males were positively associated with improvements in OSA severity (p < 0.01) but not OSA symptoms. CONCLUSION: Improvements in regional body composition were seen in males only which were related to improvements in OSA severity but not OSA symptoms. Females may exhibit different OSA pathophysiology and may require different treatment approaches. TRIAL REGISTRATION: https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=369975&isReview=trueAACTRN12616000203459 ACTRN12616000203459.

Contemporary Concise Review 2022: Sleep.

COVID-19 AND SLEEP: The COVID-19 pandemic is associated with an increase in insomnia and impaired sleep quality Health care workers are particularly susceptible and improved with cognitive behavioural therapy for insomnia (CBT-I) Long COVID has significant effects on sleep OSA impacts on the severity of acute COVID-19 illness OBSTRUCTIVE SLEEP APNOEA: Large trials of clinically representative patients confirm the cardiovascular benefits of CPAP treatment in OSA CPAP may improve long-term cognitive outcomes in OSA, but further research is needed Racial disparities in OSA prevalence and mortality risk are becoming evident Periodic evaluation of OSA risk in pregnancy is important as timing may be key for intervention to prevent or treat cardiovascular risk factors INSOMNIA: Comorbid insomnia and obstructive sleep apnoea (COMISA) can frequently co-exist and the combined negative effects of both may be deleterious, particularly to cardiovascular health There is evidence for effectiveness with novel orexin receptor antagonists.

Obstructive sleep apnea during REM sleep: effects on morning and evening blood pressure.

STUDY OBJECTIVES: Obstructive sleep apnea (OSA) is linked to the emergence and progression of cardiovascular complications including hypertension, stroke, arrhythmias, coronary artery disease, and heart failure. Epidemiological studies have reported that hypertension is associated with respiratory events during REM sleep. We examined the relationship between respiratory events during REM and morning and evening hypertensive blood pressure (BP) levels in a clinical sleep population. METHODS: This study included data from in-laboratory diagnostic polysomnographic studies (n = 797) from adults attending for investigation of OSA. Hypertensive BP levels were defined using BP measurements taken in the evening before and morning after polysomnography, and the use of antihypertensive medication. Regression modeling was undertaken to examine the probability of evening and morning hypertensive BP levels according to REM apnea-hypopnea index (AHI), NREM AHI, gender, age, body mass index (BMI), alcohol use, total sleep time (TST), sleep time SpO2 <90%, and smoking status. RESULTS: The probability of morning hypertensive BP levels was significantly independently associated with age (p < .001), BMI (p < .001), and REM AHI (p < .001). No significant effect was found for the male gender, NREM AHI, alcohol use, TST, sleep time SpO2 <90%, or smoking (p > .05 for all). The probability of evening hypertensive BP levels was only significantly associated with age (p < .001), male gender (p = .012), BMI (p < .001), and TST (p = .032). CONCLUSIONS: Respiratory events during REM sleep are significantly associated with morning hypertensive BP levels. Future research is needed to determine whether treatment of these events can prevent or reverse morning hypertensive BP levels.

A 12-month weight loss intervention in adults with obstructive sleep apnoea: is timing important? A step wedge randomised trial.

BACKGROUND/OBJECTIVES: Continuous positive airway pressure (CPAP) concomitant with weight loss is a recommended treatment approach for adults with moderate-severe obstructive sleep apnoea (OSA) and obesity. This requires multiple synchronous behaviour changes. The aim of this study was to examine the effectiveness of a 6-month lifestyle intervention and to determine whether the timing of starting a weight loss attempt affects weight change and trajectory after 12 months in adults newly diagnosed with moderate-severe OSA and treated at home with overnight CPAP. METHODS: Using a stepped-wedge design, participants were randomised to commence a six-month lifestyle intervention between one and six-months post-enrolment, with a 12-month overall follow-up. Adults (n = 60, 75% males, mean age 49.4 SD 10.74 years) newly diagnosed with moderate-severe OSA and above a healthy weight (mean BMI 34.1 SD 4.8) were recruited. RESULTS: After 12 months, exposure to the intervention (CPAP and lifestyle) resulted in a 3.7 (95% CI: 2.6 to 4.8, p < 0.001) kg loss of weight compared to the control condition (CPAP alone). Timing of the weight loss attempt made no difference to outcomes at 12 months. When exposed to CPAP only (control period) there was no change in body weight (Coef, [95% CI] 0.03, [-0.3 to 0.36], p = 0.86). CONCLUSIONS: The lifestyle intervention resulted in a modest reduction in body weight, while timing of commencement did not impact the degree of weight loss at 12 months. These findings support the recommendation of adjunctive weight-loss interventions within six-months of starting CPAP.

Effect of shift work on frontline doctors' mood and sleep during the COVID-19 pandemic.

Purpose: Shift work has detrimental effects on healthcare workers, which may be further compounded by frontline work during the COVID-19 pandemic. We postulated that sleep would worsen and distress would increase during COVID-ward service. Methods: Doctors (n = 18) were recruited from a tertiary centre during the second wave of the COVID-19 pandemic in Melbourne, Australia. Participants had been rostered ON to consecutive 7 day or night shifts and a week OFF over a fortnight. 9 worked on COVID wards managing positive/suspected COVID patients, and 9 were allocated to general MEDICAL wards. Participants wore wrist actigraphy, and completed the Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS), and Kessler Psychological Distress Scale (K10) at baseline and the end of each week. Results: Both the COVID and MEDICAL groups spent less time in bed and had reduced total sleep time during their week ON shift compared to week OFF shift. The COVID group had worse sleep quality (PSQI Δ + 1.0, 6.8 vs 5.8, p = 0.036), daytime sleepiness (ESS Δ + 2.6, 8 vs 5.4, p = 0.014) and greater distress (K10 Δ + 1.7, 17 vs 15.3, p = 0.002) during their week ON compared to BASELINE. Conclusion: During the COVID-19 pandemic shift workers had poorer sleep during their week ON. Those working on COVID wards had greater distress during their week ON than those working on general MEDICAL wards. It is important to recognise the potential for sleep deficits and greater distress in medical workers during the pandemic.

Barriers to sleep in acute hospital settings.

OBJECTIVE: This study aimed to examine the environmental and operational factors that disrupt sleep in the acute, non-ICU hospital setting. DESIGN, SETTING AND PARTICIPANTS: This was a prospective study of adult patients admitted to an acute tertiary hospital ward (shared versus single room) and sleep laboratory (single room conducive to sleep). MAIN OUTCOME MEASURES: This study measured ambient light (lux) and sound (dB), number of operational interruptions, and questionnaires assessing sleep and mental health. RESULTS: Sixty patients were enrolled, 20 in a double bedroom located close to the nursing station ('shared ward'), 20 in a single bedroom located distant to the nursing station ('single ward') and 20 attending the sleep laboratory for overnight polysomnography ('sleep laboratory'). Sleep was disturbed in 45% of patients in the shared and single ward groups (Pittsburgh Sleep Quality Index > 5). Light levels were appropriately low across all 3 locations. Sound levels (significant effect of room F(1.38) = 6.452, p = 0.015) and operational interruptions (shared ward 5.6 ± 2.5, single ward 6.2 ± 2.9, sleep laboratory 2.7 ± 2.1 per night, p < 0.05 wards compared to sleep laboratory) were higher in the shared and single ward group compared to the sleep laboratory but not compared to each other. Noise was rated as the greatest environmental disturbance by 70% of ward patients compared to 10% in the sleep laboratory. CONCLUSION: Higher noise levels and frequent operational interruptions are potential barriers to sleep and recovery on an acute medical ward which are not ameliorated by being in a single bedroom located distant to the nursing station.

Evaluating mental illness-substance abuse stigmatic perceptions through education: A library-nursing initiative.

In this library-nursing initiative, the goal was to identify any changes in stigmatic perceptions among public library staff regarding mental health and substance abuse, post 3-day educational sessions. A total of n = 37 library staff participated in this project and attended all three educational sessions and completed a pre- and postquestionnaire designed to address common mental health and substance abuse-related stigmatic perceptions among public library staff. Upon analysis of the pre- and postresponses, we identified that there were statistically significant changes in two perceptions, namely; "embarrassed to disclose mental illness" and "commonality of substance abuse" (p ≤ .05). Therefore, it is essential that behavioral health nurses engage and collaborate with local public libraries to educate and build a safe environment for vulnerable populations like those affected with mental illness and substance abuse in the community settings.

Microsleep assessment enhances interpretation of the Maintenance of Wakefulness Test.

STUDY OBJECTIVES: The Maintenance of Wakefulness Test (MWT) is used to objectively evaluate an individual's ability to remain awake; however, microsleeps are not included in the assessment. We aimed to determine if microsleep data prior to sleep onset assisted in interpretation of ability to maintain wakefulness across a range of typical patient groups. METHODS: Forty-eight patients referred for overnight polysomnography and subsequent MWT were included. Patients were divided into 3 groups (treated obstructive sleep apnea [OSA], untreated OSA, or treated idiopathic hypersomnia or narcolepsy) based on prior medical diagnosis. Demographics, clinical characteristics, polysomnography, and MWT variables, including frequency, distribution, duration, and latency of microsleeps were compared between groups. RESULTS: Microsleeps were observed in MWT trials significantly more frequently in patients with treated idiopathic hypersomnia/narcolepsy over the course of the day (0.34 ± 0.06 vs 0.07 ± 0.02 microsleeps/min; P < .001) and in patients with untreated OSA toward the end of the day (0.31 ± 0.06 vs 0.05 ± 0.02 microsleeps/min; P < .001) compared to the group with treated OSA. Microsleeps were often observed in series and earlier in patients with treated idiopathic hypersomnia/narcolepsy (10.9 ± 1.6 minutes) and those with untreated OSA (16.2 ± 2.7 minutes) compared to the group with treated OSA (24.9 ± 3.0 minutes; P < .05), and, if taken into consideration, would increase the proportion of patients demonstrating inability to maintain wakefulness by 33% and 22%, respectively. CONCLUSIONS: MWT performance varies significantly across patient groups. Microsleep analysis prior to sleep onset may be a more sensitive measure of patient daytime wakefulness than sleep latency alone and should be considered in MWT assessment. CITATION: Anniss AM, Young A, O'Driscoll DM. Microsleep assessment enhances interpretation of the Maintenance of Wakefulness Test. J Clin Sleep Med. 2021;17(8):1571-1578.

Psychoactive drug and medication use among patients referred to a tertiary sleep laboratory population.

Psychoactive drugs including alcohol, caffeine, and prescription medications are commonly consumed to alter sleep/wake states, however the prevalence and impact of these drugs among populations seeking assessment from sleep physicians are unknown. We investigated the prevalence of commonly used drugs (alcohol and caffeine), and medications in a population (N=120; 50 females and 70 males) attending a tertiary sleep clinic for diagnostic polysomnography (PSG) assessment. In addition to objective sleep assessment, participants completed questionnaires assessing sleep quality (Pittsburgh Sleep Quality Index, PSQI), daytime sleepiness (Epworth Sleepiness Scale, ESS), depression and anxiety (Hospital Anxiety and Depression Scale, HADS), alcohol use (Alcohol Use Disorders Identification Test, AUDIT), caffeine and medication use, as well as their experience of adverse events (motor vehicle accidents and near-miss crashes). Caffeine was consumed by 90% of the population and was associated with a reduction in excessive sleepiness symptomology; while high AUDIT scores were associated with increased near-miss crashes. Polypharmacy was common, with a greater number of medications associated with poorer sleep quality, and changes in sleep architecture. This study maps commonly used drugs in those attending a tertiary sleep clinic, and demonstrates associations between drug use and sleep outcomes assessed objectively and subjectively.

Making Fall Risk Assessment Clinically Relevant in an Adult Psychiatric Setting.

The purpose of this non-experimental descriptive study was to measure psychiatric clinical nurses' (N = 25) perceptions of the Edmonson Psychiatric Falls Risk Assessment Tool© (EPFRAT) compared to the Morse Fall Scale (MFS) and to evaluate patient falls with injury rates 12 months before and after the study. The setting was a 27-bed, adult psychiatric unit in a community-based teaching hospital located in the Northeast region of the United States. The EPFRAT and MFS were used to assess fall risk in 216 patients over 3 months. Findings indicated that the EPFRAT was more user-friendly and relevant; improved nurses' clinical judgment in identifying high-risk patients; and nurses were supportive toward changing practice from using the MFS to EPFRAT for fall risk assessment. Falls with injury rates decreased by 0.52 per 1,000 patient care days post-implementation of the EPFRAT. [Journal of Psychosocial Nursing and Mental Health Services, 58(2), 21-26.].

Effects of Animal-Assisted Activity on Mood States and Feelings in a Psychiatric Setting.

BACKGROUND: Research has shown that animal-assisted activity (AAA) effectively improves physiological, psychological, emotional, and social well-being in various environments. AIMS: To identify how AAA affects mood states and feelings among both patients and staff on inpatient psychiatric units. METHODS: This study used a quasi-experimental, pre-/posttest design with nonequivalent comparison groups. Quantitative data were collected using the Visual Analog Mood Scale. Demographic data, with an open-ended question, were obtained postsessions. RESULTS: Significant changes were observed pre- and postexposure to AAA sessions with a therapy dog. Negative moods decreased, and positive moods increased as measured by the Visual Analog Mood Scale. Content analysis identified themes of feeling happy, feeling relaxed, and feeling calm. CONCLUSIONS: This research expands scientific evidence associated with AAA by identifying changes in mood states and feelings among individuals in a psychiatric setting.

The physiological phenotype of obstructive sleep apnea differs between Caucasian and Chinese patients.

STUDY OBJECTIVES: The mechanisms responsible for the development of obstructive sleep apnea (phenotypic "traits") are known to differ between individuals and may differ across ethnicities. We aimed to examine whether loop gain, arousal threshold, pharyngeal collapsibility and muscle compensation differ between Chinese and Caucasian individuals with OSA. METHODS: We noninvasively determined the relative contribution of loop gain, arousal threshold, pharyngeal collapsibility, and muscle compensation from the ventilatory flow pattern recorded during a standard clinical polysomnography in a cohort of age and AHI matched Caucasian and Chinese patients with moderate-severe OSA (n = 90). RESULTS: Chinese participants had significantly more collapsible pharyngeal airways as indicated by a lower Vpassive (68.9 [51.5-75.2] vs. 74.0 [65.1-80.4] %Veupnea, U = 703, p = 0.012), but less ventilatory control instability as indicated by a lower loop gain (0.60 [0.50-0.67] vs. 0.63 [0.57-0.81], U = 762, p = 0.043) compared with Caucasian participants. Further, multiple logistic regression analyses demonstrated that the combined pharyngeal collapsibility (Vpassive) and loop gain traits help to better explain the differences between the groups beyond upper-airway collapsibility alone. No statistically significant group differences were found in muscle compensation or arousal threshold traits between groups. CONCLUSION: Individuals of Chinese descent appear to have OSA that is driven much more by the relative contribution of their anatomical predisposition and to a lesser extent nonanatomical causes compared with Caucasians. Future research should focus on determining if Chinese versus Caucasian ethnicity is an important contributing factor to clinical outcomes and therapeutic responses in OSA.

Assessing the impact of diet, exercise and the combination of the two as a treatment for OSA: A systematic review and meta-analysis.

This study aimed to provide an updated systematic review and meta-analysis of randomized controlled trials (RCT) investigating the effectiveness of lifestyle interventions on weight loss and the impact on the severity of obstructive sleep apnoea (OSA). A systematic search of five databases between 1980 and May 2018 was used to identify all RCT which employed a lifestyle intervention (i.e. diet-only, exercise-only or combination of the two) aiming to reduce the severity of OSA (assessed using the apnoea-hypopnoea index (AHI)). Random-effects meta-analyses followed by meta-regression were conducted. Ten RCT involving 702 participants (Intervention group: n = 354; Control group: n = 348) were assessed in two meta-analyses. The weighted mean difference in AHI (-8.09 events/h, 95% CI: -11.94 to -4.25) and body mass index (BMI, -2.41 kg/m2 , 95% CI: -4.09 to -0.73) both significantly favoured lifestyle interventions over control arms. Subgroup analyses demonstrated that all interventions were associated with reductions in the AHI, but only the diet-only interventions were associated with a significant reduction in BMI. No association was found between the reduction in AHI or BMI and the length of the intervention, or with baseline AHI and BMI levels. All lifestyle interventions investigated appear effective for improving OSA severity and should be an essential component of treatment for OSA. Future research should be directed towards identifying subgroups likely to reap greater treatment benefits as well as other therapeutic benefits provided by these interventions.

Sleeping Well Trial: Increasing the effectiveness of treatment with continuous positive airway pressure using a weight management program in overweight adults with obstructive sleep apnoea-A stepped wedge randomised trial protocol.

BACKGROUND: The majority of adults diagnosed with obstructive sleep apnoea (OSA) are overweight or obese. Continuous positive airway pressure (CPAP) is the most common effective therapy for OSA. However, adherence declines over time with only 50% of patients prescribed CPAP continuing to use it long term. Furthermore, a recent prospective analysis indicated that those more adherent with CPAP therapy have enhanced weight gain trajectories which in turn may negatively impact their OSA. AIM: The Sleeping Well Trial aims to establish whether the timing of starting a lifestyle weight loss intervention impacts on weight trajectory in those with moderate-severe OSA treated at home with CPAP, while testing the potential for smart phone technology to improve adherence with lifestyle interventions. METHODS: A stepped wedge design with randomisation of individuals from 1 to 6 months post-enrolment, with 5 months of additional prospective follow up after completion of the stepped wedge. This design will investigate the effect of the 6-month lifestyle intervention on people undergoing CPAP on body weight, body composition and health-related quality of life. DISCUSSION: This trial tests whether the timing of supporting the patient through a weight loss intervention is important in obtaining the maximum benefit of a lifestyle change and CPAP usage, and identify how best to support patients through this critical period. TRIAL REGISTRATION: The protocol (v1) is registered prospectively with the International Clinical Trials Registry (CTR) ACTRN12616000203459 (public access). Any amendments to protocol will be documented via the CTR. Recruitment commenced in March 2016 with data collection scheduled to finish by May 2018.

The use of accelerometry as a tool to measure disturbed nocturnal sleep in Parkinson's disease.

Sleep disturbances are common in Parkinson's disease (PD). We used the Parkinson's KinetiGraph (PKG), an objective movement recording system for PD to assess night time sleep in 155 people aged over 60 and without PD (controls), 72 people with PD (PwP) and 46 subjects undergoing a Polysomnogram (PSG: 36 with sleep disorder and 10 with normal sleep). The PKG system uses a wrist worn logger to capture acceleration and derive a bradykinesia score (BKS) every 2 min over 6 days. The BKS ranges from 0-160 with higher scores associated with lesser mobility. Previously we showed that BKS > 80 were associated with day time sleep and used this to produce scores for night time sleep: Efficiency (Percent time with BKS > 80), Fragmentation (Average duration of runs of BKS > 80) and Sleep Quality (BKS > 111 as a representation of atonia). There was a fair association with BKS score and sleep level as judged by PSG. Using these PKG scores, it was possible to distinguish between normal and abnormal PSG studies with good Selectivity (86%) and Sensitivity (80%). The PKG's sleep scores were significantly different in PD and Controls and correlated with a subject's self-assessment (PDSS 2) of the quality, wakefulness and restlessness. Using both the PDSS 2 and the PKG, it was apparent that sleep disturbances were apparent early in disease in many PD subjects and that subjects with poor night time sleep were more likely to have day time sleepiness. This system shows promise as a quantitative score for assessing sleep in Parkinson's disease.

Importance of Urinary Drug Screening in the Multiple Sleep Latency Test and Maintenance of Wakefulness Test.

STUDY OBJECTIVES: Multiple sleep latency testing (MSLT) and the maintenance of wakefulness test (MWT) are gold-standard objective tests of daytime sleepiness and alertness; however, there is marked variability in their interpretation and practice. This study aimed to determine the incidence of positive drug screens and their influence on MSLT, MWT, and polysomnographic variables. METHODS: All patients attending Eastern Health Sleep Laboratory for MSLT or MWT over a 21-mo period were included in the study. Urinary drug screening for amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine, methadone, and opiates was performed following overnight polysomnography (PSG). Demographics and PSG variables were compared. RESULTS: Of 69 studies, MSLT (43) and MWT (26), 16% of patients had positive urinary drug screening (7 MSLT; 4 MWT). Drugs detected included amphetamines, cannabinoids, opiates, and benzodiazepines. No patient self-reported use of these medications prior to testing. No demographic, MSLT or MWT PSG data or overnight PSG data showed any statistical differences between positive and negative drug screen groups. Of seven MSLT patients testing positive for drug use, one met criteria for the diagnosis of narcolepsy and five for idiopathic hypersomnia. On MWT, three of the four drug-positive patients had a history of a motor vehicle accident and two patients were occupational drivers. CONCLUSIONS: These findings indicate drug use is present in patients attending for daytime testing of objective sleepiness and wakefulness. These data support routine urinary drug screening in all patients undergoing MSLT or MWT studies to ensure accurate interpretation in the context of illicit and prescription drug use.

Overnight Motor Skill Learning Outcomes in Obstructive Sleep Apnea: Effect of Continuous Positive Airway Pressure.

STUDY OBJECTIVE: To determine the effectiveness of continuous positive airway pressure (CPAP) therapy in alleviating known impairments in the overnight consolidation of motor skill learning in patients with obstructive sleep apnea (OSA). METHODS: Twenty-five patients with untreated moderate-severe OSA, 13 first-night CPAP users, 17 compliant CPAP users, and 14 healthy control patients were trained on a motor sequence learning task (Sequential Finger Tapping Task, SFTT) and were subsequently tested prior to and after polysomnographic recorded sleep. Measures of subjective sleepiness (Karolinska Sleepiness Scale) and sustained attention (Psychomotor Vigilance Task) were also completed before and after sleep. RESULTS: Typical analyses of overnight improvement on the SFTT show significantly greater overnight gains in motor task speed in controls (+11.6 ± 4.7%, p = 0.007) and compliant CPAP users (+8.9 ± 4.3%, p = 0.008) compared to patients with OSA (-4.86 ± 4.5%). Additional analyses suggest that these improvements in motor performance occurred prior to the sleep episode, as all groups significantly improved (15% to 22%) over a 10-min presleep rest period. Thereafter, performance in all groups significantly deteriorated over sleep (6% to 16%) with trends toward patients with OSA showing greater losses in performance compared to control patients and compliant CPAP users. No between-group differences in subjective sleepiness and sustained attention were found presleep and postsleep. CONCLUSIONS: The current data suggest impairments in overnight motor learning in patients with OSA may be a combination of deficient stabilization of memory over a sleep episode as well as increased vulnerability to time on task fatigue effects. Compliant CPAP usage possibly offsets both of these impediments to learning outcomes by improving both sleep quality and subsequent daytime function.

College of American Pathologists' laboratory standards for next-generation sequencing clinical tests.

CONTEXT: The higher throughput and lower per-base cost of next-generation sequencing (NGS) as compared to Sanger sequencing has led to its rapid adoption in clinical testing. The number of laboratories offering NGS-based tests has also grown considerably in the past few years, despite the fact that specific Clinical Laboratory Improvement Amendments of 1988/College of American Pathologists (CAP) laboratory standards had not yet been developed to regulate this technology. OBJECTIVE: To develop a checklist for clinical testing using NGS technology that sets standards for the analytic wet bench process and for bioinformatics or "dry bench" analyses. As NGS-based clinical tests are new to diagnostic testing and are of much greater complexity than traditional Sanger sequencing-based tests, there is an urgent need to develop new regulatory standards for laboratories offering these tests. DESIGN: To develop the necessary regulatory framework for NGS and to facilitate appropriate adoption of this technology for clinical testing, CAP formed a committee in 2011, the NGS Work Group, to deliberate upon the contents to be included in the checklist. Results . -A total of 18 laboratory accreditation checklist requirements for the analytic wet bench process and bioinformatics analysis processes have been included within CAP's molecular pathology checklist (MOL). CONCLUSIONS: This report describes the important issues considered by the CAP committee during the development of the new checklist requirements, which address documentation, validation, quality assurance, confirmatory testing, exception logs, monitoring of upgrades, variant interpretation and reporting, incidental findings, data storage, version traceability, and data transfer confidentiality.